这里描述的工作的资金来自加利福尼亚农药管理局和加州水资源部。

城市生物威胁效能监测<ol><li> 风水取样 <ol><li>样本4 L的预处理暴雨水进入生物危机入口时离开停车场，然后通过4"出口排水口离开生物威胁后的4升后处理雨水。 </li><li>使用当地天气预报，在风暴水文图的开始，中间和结束时收集样本。在风暴事件期间复合样本以表征径流变率。 </li><li>手动收集1.3升样品，并将其复合到4升琥珀色瓶中。收集入口样本在几个路边开口，暴雨水流入生物威胁。 </li><li>从连接到出口排水管（如下所述）的流量计收集1.3L出口样品，并将其复合到4升琥珀色瓶中。 </li><li>将合成的样品储存在冰上，直到收集最终的水文样品。然后将它们运送到实验室，并保持一个refr在4℃下进行化学和毒性测试的二次采样之前。在样品采集48小时内将样品送至化学实验室。 </li></ol></li><li> 负荷计算 <ol><li>在暴风雨之前，安装一个倾倒式数字记录仪雨量计，将其连接到与bioswale现场相邻的灯或其他杆。使用雨数据来表明现场的瞬时和总降雨量。 </li><li>在生物威胁的出口排水管上安装机械齿轮脉冲流量计。记录从bioswale出来的总流量。 注意:假设减少径流量可以减少LID设计中污染物的总体负荷。 </li><li>使用雨量记录下来的英寸雨量，通过外推模拟雨水期间落在停车场集水区的水量。使用这些数据确定基于停车场表面积进入处理系统的体积。 </li><li>使用t记录的总流量他出口流量计计算渗透百分比。计算入口和出口体积之间的差异，以确定雨水渗透。 </li><li>使用入口和出口体积结合污染物分析测量计算风暴期间的污染物负荷和负荷减少百分比。 </li><li>测量与地表水毒性相关的化学分析物（如下所述）。总化学基团简化负荷计算，并基于其相似的毒性作用模式（ 如总多环芳烃[PAHs]，总拟除虫菊酯和总氟虫腈和降解物）。 </li></ol></li><li> 化学 <ol><li>分析以下参数的所有样品:总悬浮固体（TSS），痕量金属（USEPA方法200.8 5 ;电感耦合等离子体质谱[ICP / MS]）和PAHs（USEPA方法625 6 ）。 </li><li>分析样品包括9种拟除虫菊酯（USEPA方法SW846 8270改性7 ;联苯菊酯，氯氰菊酯，氰戊菊酯/氰戊菊酯，氯菊酯，溴氰菊酯，氯氰菊酯，氟氯氰菊酯和烯丙菊酯），氟虫腈及其三种主要降解物（氟虫腈，氟虫腈砜，氟虫腈脱亚硫酰基）。 </li><li>使用气相色谱 - 质谱（GC / MS），使用负电化学电离或其他合适的方法分析拟除虫菊酯以提供足够的检测限。由于大多数目前使用的农药在低浓度下具有高毒性，因此其分析要求低的化学报告限值与环境风险评估相关。所有除虫菊酯除外，拟除虫菊酯的方法报告限为0.5 ng / L至1.0 ng / L（报告限为10 ng / L）。 </li><li>使用氟虫腈的分析程序，提供1.0 ng / L的方法报告限值。有机磷农药不需要测量在当地使用模式，例如在加利福尼亚州的市区8,9 。 </li><li>使用与三重四极杆质谱联用的超高效液相色谱法测量新烟碱类农药（ 如吡虫啉），其具有50ng / L的吡虫啉报告限。 </li></ol></li><li> 毒性测试 <ol><li>根据美国环境保护局（USEPA）急性试验方案，对3种试验物种进行复合入口和出口雨水样品的毒性试验10 。 96岁以上的桡足类痢疾试验测定存活率。测试与amphipod Hyalella azteca测量10天后的生存。用枸杞子（Chironomus dilutus）测试10天后的生存和生长。 </li><li>进行急性96小时生存测试与U.美国环保局指导。 <ol><li>在入口和出口雨水样本的五个重复的每一个中暴露五个C.dubia新生儿。复制品由含有15 mL测试溶液的20 mL闪烁瓶组成。 </li><li>在每天100％更新雨水测试解决方案之前，饲料将新生酵母，叶绿素，鳟鱼（= YCT，遵循美国环保署指导）和Selenastrum藻类的混合物新鲜。每天记录幸存新生儿的总数。 </li><li>比较使用t检验在96小时暴露于入口和出口暴雨水样品后，在中度硬对照水中生存的最终C.dubia存活。遵循美国环保局推荐的统计程序。 </li></ol></li><li>根据美国环保署的指导，与amphipod H. azteca进行急性10 d生存测试。 <ol><li>在五个重复的每一个中暴露10,9天到15天的两栖动物。复制品由含有200 mL测试溶液的300 mL玻璃烧杯组成。 </li><li&gt;进行两栖动物试验10天，计算每日生存的两栖动物数量，每48小时更新50％的测试溶液。每48小时喂一次烧杯，1.5升YCT更新后。 </li><li>将雨水样品中两栖动物的最终存活率与上述实验室水中的10天生存率进行比较。 </li></ol></li><li>根据美国环保局的指导，与idge藜进行慢性10 d生存和生长试验。 <ol><li>在四个重复的每一个中暴露12,7 d的老动物。复制品由含有200 mL测试溶液的300 mL玻璃烧杯组成。为每个Midge测试容器提供5 mL沙子作为幼体管建造的基底。 </li><li>对每个烧杯每48小时进行一次10天的试验，每日更换一次50％的试验溶液，每日一次，随着鱼粉的浓度越来越高（4g / L），第0至3天为0.5mL /天。第4至6天，1.0mL /天;天7至10，1.5 mL /天。 </li><li>比较最终的生存和如上所述，在实验室井水中将雨水样品的生长增加到10天生存期。测量存活动物的生长与无测试生物体的初始重量相比在10天时无灰分干燥。 </li></ol></li><li>对于所有毒性测试，使用适当的仪表和电极测量溶解氧，pH和电导率。使用分光光度计测量未离子化的氨。 <ol><li>在测试开始和结束时测量水硬度和碱度。 10 </li><li>用连续记录温度计记录水温。 </li></ol></li></ol></li></ol>综合植被农业排水沟功效监测<ol><li> 综合沟施工 注:本例中使用的农用排水沟长152米，顶部半米宽为5米，深1米。沟渠植被是n的组合主要种植有红羊茅（ Festuca rubra ）的草本植物。在这个例子中，综合营养沟试验由颗粒状活性炭（GAC）和与植被沟结合的堆肥过滤处理组成。 <ol><li>构建两个堆肥过滤器和六个碳过滤器，并将其安装在植被沟的三个不同部分（ 图2 ）。使用2米长20厘米直径的袖子，填充碳或堆肥。 </li><li>填充六个袖子与30 L的颗粒活性炭，并将它们放置在146米点的沟渠，靠近152米植被沟的尽头。将GAC填充的套管锚定到沟槽底部，并在上游边缘的钢丝绳上。 </li><li>在每个GAC套管的下游边缘放置一个2.5米长的6"宽松松板，将松木板松开通道的两侧和底部，以最大限度地减少水旁路和​​切割碳套筒，板也将亲提供垂直支持，以最大限度地提高与碳的水接触时间。 </li><li>在堆肥套筒上填充约15公斤的每个部分分解的废弃物，从任何干净的来源，如当地垃圾填埋场。将两条长2米长的堆肥套筒沿着植被沟渠放置，长度为64米，距离152米植被沟的长度为123米（ 图2 ）。 </li></ol></li><li> 径流模拟和抽样 注意:本协议描述了进行模拟农业径流试验和相关监测的方法，以使用综合营养处理系统评估治疗效果。在当前的例子中，综合植被堆肥碳体系以两个流速进行了评估，分别代表萨利纳斯山谷商业农场的典型场外排放量，3.2升/秒和6.3升/秒。有机磷农药毒死蜱在这些试验中被用作模型农药，因为它具有适度的溶解度因此代表了常用于病虫害管理的代表性农药的中等程度的溶解度。毒死蜱也是加州中部正在进行的监管行动的主题，因为它对农业流域的影响。目标毒死蜱剂量约为2,600ng / L。流量和目标毒死蜱浓度在以前在当地灌溉径流3,11中测量的范围内。在这里给出的例子中，没有监测到经过植被沟的水脉冲的水力停留时间。在这些系统中的停留时间随着水的流入速率，先前的灌溉和下雨引起的土壤饱和度的程度，堰坝和沉积盆地的阻塞结构的存在以及被植被覆盖的表面积的数量而变化。以前的研究表明，小型沟渠系统的停留时间为数小时萨利纳斯山谷3，4 。视觉观察表明，GAC过滤器的停留时间为1或2分钟。 <ol><li>使用与悬浮泥沙混合的地下水建立模拟农业径流。对于使用模型农药毒死蜱的试验，通过将已认证的储备溶液加入已知体积的蒸馏水中，每3.2L / s试验制备10mg / L的新鲜储备溶液。每6.3L / s试验准备20mg / L的新鲜毒死蜱储备溶液。 <ol><li>在进入植被处理沟渠入口之前，使用计量泵向径流水提供一致的储备溶液体积。使用计量泵以50毫升/分钟的速度送入模拟灌溉水的流量。 </li></ol></li><li>使用数字仪表监测入口流量，并使用这些数据量化施加到沟渠入口的径流水总量。 </li><li>在呃建造堰沟槽的出口和铅垂，并连接到数字流量计的出口管。使用此仪表记录离开沟渠的径流量。 </li><li>使用连接到数字仪表的数据记录器以5分钟的间隔记录流量。编制数据记录器以激活位于入口处和沟渠入口下方各个站（ 例如 ，23 m，45 m和68 m）的蠕动泵，以5分钟间隔收集不锈钢容器的复合子样品。 </li></ol></li><li> 化学 <ol><li>在每个径流试验结束时，将试验中的径流水复合样品转移到琥珀色玻璃瓶中，并将样品保持在4℃的冰上，以进行毒性和化学分析。 </li><li>使用GC-MS或酶联免疫吸附试验（ELISA）分析总悬浮固体（TSS）和毒死蜱的复合材料样品。 </li><li>将"入口"复合样品（预处理）与＃＃34;出口"复合样品（后处理），以评估综合沟渠系统降低TSS和农药负荷的功效。 </li></ol></li><li> 毒性测试 <ol><li>在每次试验的入口（预处理）和出口（后处理）的复合样品中，使用96小时茜草nia毒毒性试验10确定水柱毒性，如上所述用于生物威胁监测。 C. dubia是对毒死蜱的敏感性（中位数致死浓度（LC50）= 53ng / L 12 ）的农业径流毒性的适当监测物种。 </li></ol></li></ol>

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作者宣称他们没有竞争的经济利益。

本议定书中描述的做法旨在作为消除农业灌溉和雨水径流污染物总体战略的最后步骤。使用bioswales和其他城市绿色基础设施LID做法旨在作为解决径流污染物到达相邻接收水域的最后一个难题。该协议强调监测城市生物风险的方法，以确定消除与城市污染物相关的毒性的治疗效果，重点是当前使用的农药。 设计监测研究的关键步骤包括建模方法和采样设计，以捕获暴风雨水文?...

地表水毒性在加利福尼亚流域普遍存在，几十年的监测表明毒性往往是由于农药和其他污染物1 。地表水污染的主要来源是城市和农业来源的雨水和灌溉径流。由于污染物被列为降解的污染物，并且从城市和农业来源鉴定出毒性，水质监管机构与国家和联邦资金来源合作，实施减少污染物负荷的做法。加州城市流域正在推广绿色基础设施，以减少洪水，并通过渗透和储存增加雨水的恢复。虽然低影响力开发（LID）设计被授权在许多地区进行新的建设，但很少有研究监测这些系统的功效，超出常规污染物的测量，如溶解固体，金属和液化气丝宝集团。更密集的监测最近已经评估了降低表面水毒性的化学浓度和化学负荷，并直接确定生物燃料游戏是否减少了径流的毒性。这表明bioswales有效地消除了与一些污染物类别2有关的毒性，但需要对新兴化学品进行额外的研究。 在加利福尼亚的农业流域也正在实施植被处理系统，这些已被证明可以有效地减少农业灌溉径流中的农药和其他污染物3,4 。这些系统代表了一套方法的组成部分，以减少污染物对地表水的负荷。因为它们旨在减轻负责地表水毒性的污染物，所以实施过程的关键组成部分是监测e确保其长期有效性。监测包括对化学品的化学分析以及敏感指标物种的毒性测试。本文介绍了城市停车场bioswale和农业植被排水沟系统的协议和监测结果。 典型的停车场生物威胁的设计属性，例如可用于治理典型的混合型城市购物区的风暴径流，取决于被处理的区域。在这里描述的例子中，53,286平方英尺的沥青产生了一个不透水的表面积，排水到一个洼地，由4,683平方英尺的美化组成。为了适应这个表面积的径流，一个215英尺长的平底半V形通道包括斜坡小于50％，纵向斜率为1％的洼地（ 图1 ）。这种麝香草包括三层，包括种植在6英寸表层土中的天然束草红色超过2.5英尺的压实路基。雨水从停车场流向沿着洼地的多个入口点。水浸入植被区域，然后渗入路基并排入4英寸穿孔排水沟。这个系统通过系统排水到相邻的湿地，最终排入当地的小溪。

<table border="0" cellpadding="0" cellspacing="0" width="679">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">HOBO tipping-bucket digital logger rain gauge&#160;</td>
			<td style="width:177px;">Onset Computer Co., Bourne MA, USA)</td>
			<td style="width:119px;">Onset RG3</td>
			<td style="width:167px;">Rain gauge</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Mechanical geared pulse flow meter&#160;</td>
			<td style="width:177px;">Seametrics Inc., Kent WA</td>
			<td>Seametrics MJ-R</td>
			<td>Flow meter for measuring bioswale outlet flow</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Filtrexx SafteySoxx</td>
			<td style="width:177px;">Filtrexx Co. - info@filtrexx.com</td>
			<td style="width:119px;">SafetySoxx</td>
			<td>perforated synthetic cloth for granulated activated carbon and compost</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Granulated activated carbon&#160;</td>
			<td style="width:177px;">Evoqua - Siemens Corp., Oakland CA</td>
			<td style="width:119px;">AC380</td>
			<td>GAC for agriculture irrigation water treatment</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Digital flow meters&#160;</td>
			<td style="width:177px;">Seametrics Inc. Kent WA</td>
			<td style="width:119px;">Ag2000; WMP101</td>
			<td>Flow meters for agriculture irrigation treatment system monitoring</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Data Loggers</td>
			<td>Campbell Scientific Inc., Logan, UT</td>
			<td style="width:119px;">CR1000</td>
			<td>Data loggers for recording flow data</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Peristaltic pumps for composite sampling</td>
			<td style="width:177px;">Omega Engineering Inc. Stamford CT</td>
			<td>Omegaflex FPU-122-12VDC&#160;</td>
			<td>Pumps for composite sampling</td>
		</tr>
	</tbody>
</table>

vegetated treatment systems for removing contaminants associated with surface water toxicity in agriculture and urban runoff

城市雨水和农业灌溉径流含有对相邻接收水经常有毒的污染物的复杂混合物。径流可以通过设计用于促进污染物吸附到植被和土壤并促进渗透的简单系统进行处理。描述了两个示例系统:城市雨水处理的生物威胁处理系统和用于处理农业灌溉径流的植被排水沟。两者具有类似的属性，可以减少径流中的污染物负荷:导致污染物吸收到土壤和植物表面的植被以及水渗透。这些系统还可以包括作为抛光步骤的颗粒状活性炭的整合以去除残留的污染物。这些系统在农业和城市流域的实施需要系统监测来验证治疗效果。这包括对负责毒性的特定污染物进行化学监测。本文强调监测目前使用的农药，因为这些农药对水生无脊椎动物造成地表水的毒性。

城市生物威胁效能在风暴18.5小时内，雨量计记录下来的雨量为1.52"，这导致了从停车场流入生物圈的水流量为50,490加仑，其中出口流量计记录了5248加仑，导致90％的雨水流入生物威胁，生物威胁减少了所有化学品的监测，总悬浮固体减少了72％（ 表1 ），检测到多环芳烃浓度非常低，但所有浓度的多环芳烃降低100％?...

Watch this Scientific Journal Video about Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff at JoVE.com

Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

本文总结了处理城市雨水和农业灌溉径流去除与水生毒性相关的农药和其他污染物的处理系统的设计属性和有效性。

用于去除与农业和城市径流中地表水毒性有关的污染物的植被处理系统

Urban stormwater and agriculture irrigation runoff contain a complex mixture of contaminants that are often toxic to adjacent receiving waters. Runoff may ...

vegetated-treatment-systems-for-removing-contaminants-associated-with

Research

JoVE Journal

Environment

10.5K 次观看.  University of California, Davis. 本文总结了处理城市雨水和农业灌溉径流去除与水生毒性相关的农药和其他污染物的处理系统的设计属性和有效性。 

视频: Vegetated Treatment Systems for Removing Contaminants Associated with Surface Water Toxicity in Agriculture and Urban Runoff

Design and Construction of an Urban Runoff Research Facility

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use due to increased demand for landscape irrigation. Improper irrigation practices and large rainfall events can result in runoff from urban landscapes which has potential to carry nutrients and sediments into local streams and lakes where they may contribute to eutrophication. A 1,000 m2 facility was constructed which consists of 24 individual 33.6 m2 field plots, each equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated urban landscapes. Runoff volumes from the first and second trials had coefficient of variability (CV) values of 38.2 and 28.7%, respectively. CV values for runoff pH, EC, and Na concentration for both trials were all under 10%. Concentrations of DOC, TDN, DON, PO4-P, K+, Mg2+, and Ca2+ had CV values less than 50% in both trials. Overall, the results of testing performed after sod installation at the facility indicated good uniformity between plots for runoff volumes and chemical constituents. The large plot size is sufficient to include much of the natural variability and therefore provides better simulation of urban landscape ecosystems.

This paper describes the design, construction, and function of a 1,000 m2 facility containing 24 individual 33.6 m2 field plots equipped for measuring total runoff volumes with time and collection of runoff subsamples at selected intervals for quantification of chemical constituents in the runoff water from simulated home lawns.

设计一个城市径流研究设施的建设

As the urban population increases, so does the area of irrigated urban landscape. Summer water use in urban areas can be 2-3x winter base line water use ...

科研

环境科学

A Protocol for Conducting Rainfall Simulation to Study Soil Runoff

Rainfall is a driving force for the transport of environmental contaminants from agricultural soils to surficial water bodies via surface runoff. The objective of this study was to characterize the effects of antecedent soil moisture content on the fate and transport of surface applied commercial urea, a common form of nitrogen (N) fertilizer, following a rainfall event that occurs within 24&#160;hr&#160;after fertilizer application. Although urea is assumed to be readily hydrolyzed to ammonium and therefore not often available for transport, recent studies suggest that urea can be transported from agricultural soils to coastal waters where it is implicated in harmful algal blooms. A rainfall simulator was used to apply a consistent rate of uniform rainfall across packed soil boxes that had been prewetted to different soil moisture contents. By controlling rainfall and soil physical characteristics, the effects of antecedent soil moisture on urea loss were isolated. Wetter soils exhibited shorter time from rainfall initiation to runoff initiation, greater total volume of runoff, higher urea concentrations in runoff, and greater mass loadings of urea in runoff. These results also demonstrate the importance of controlling for antecedent soil moisture content in studies designed to isolate other variables, such as soil physical or chemical characteristics, slope, soil cover, management, or rainfall characteristics. Because rainfall simulators are designed to deliver raindrops of similar size and velocity as natural rainfall, studies conducted under a standardized protocol can yield valuable data that, in turn, can be used to develop models for predicting the fate and transport of pollutants in runoff.

A rainfall simulator was used to apply a consistent rate of uniform rainfall to packed soil boxes in a study of the fate and transport of urea, a nonpoint source environmental contaminant. Under uniform soil and rainfall conditions, antecedent soil moisture content exerted strong control over urea loss in surface runoff.

一个协议，用于开展降雨模拟研究土壤径流

Rainfall is a driving force for the transport of environmental contaminants from agricultural soils to surficial water bodies via surface runoff. The ...

A Whole Cell Bioreporter Approach to Assess Transport and Bioavailability of Organic Contaminants in Water Unsaturated Systems

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is not an appropriate measure for its availability; bioavailability rather depends on the dynamic interplay of potential mass transfer (flux) of a compound to a microbial cell and the capacity of the latter to degrade the compound. In water-unsaturated parts of the soil, mycelia have been shown to overcome bioavailability limitations by actively transporting and mobilizing organic compounds over the range of centimeters. Whereas the extent of mycelia-based transport can be quantified easily by chemical means, verification of the contaminant-bioavailability to bacterial cells requires a biological method. Addressing this constraint, we chose the PAH fluorene (FLU) as a model compound and developed a water unsaturated model microcosm linking a spatially separated FLU point source and the FLU degrading bioreporter bacterium Burkholderia sartisoli RP037-mChe by a mycelial network of Pythium ultimum. Since the bioreporter expresses eGFP in response of the PAH flux to the cell, bacterial FLU exposure and degradation could be monitored directly in the microcosms via confocal laser scanning microscopy (CLSM). CLSM and image analyses revealed a significant increase of the eGFP expression in the presence of P. ultimum compared to controls without mycelia or FLU thus indicating FLU bioavailability to bacteria after mycelia-mediated transport. CLSM results were supported by chemical analyses in identical microcosms. The developed microcosm proved suitable to investigate contaminant bioavailability and to concomitantly visualize the involved bacteria-mycelial interactions.

A whole cell bioreporter assay with Burkholderia sartisoli RP037-mChe was developed to detect fractions of an organic contaminant (i.e., fluorene) available for bacterial degradation after active transport by mycelia bridging air-filled pores in a water unsaturated model system.

全细胞Bioreporter方法​​评估交通有机污染物的生物利用度和水饱和系统

Bioavailability of contaminants is a prerequisite for their effective biodegradation in soil. The average bulk concentration of a contaminant, however, is ...

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

准备和测试阻抗为基础与RTgill-W1细胞饮用水进行快速评估水样进行毒性射流生物芯片

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

This manuscript describes a duplex digital PCR assay (EntHF183 dPCR) for simultaneous quantification of Enterococcus spp. and the human fecal-associated HF183 marker. The EntHF183 duplex dPCR (referred as EntHF183 dPCR hereon) assay uses the same primer and probe sequences as its published individual quantitative PCR (qPCR) counterparts. Likewise, the same water filtration and DNA extraction procedures as performed prior to qPCR are followed prior to running dPCR. However, the duplex dPCR assay has several advantages over the qPCR assays. Most important, the dPCR assay eliminates the need for running a standard curve and hence, the associated bias and variability, by direct quantification of its targets. In addition, while duplexing (i.e. simultaneous quantification) Enterococcus and HF183 in qPCR often leads to severe underestimation of the less abundant target in a sample, dPCR provides consistent quantification of both targets, whether quantified individually or simultaneously in the same reaction. The dPCR assay is also able to tolerate PCR inhibitor concentrations that are one to two orders of magnitude higher than those tolerated by qPCR. These advantages make the EntHF183 dPCR assay particularly attractive because it simultaneously provides accurate and repeatable information on both general and human-associated fecal contamination in environmental waters without the need to run two separate qPCR assays. Despite its advantages over qPCR, the upper quantification limit of the dPCR assay with currently available instrumentation is approximately four orders of magnitude lower than that achievable by qPCR. Consequently, dilution is needed for measurement of high concentrations of target organisms such as those typically observed following sewage spills.

A Duplex Digital PCR Assay for Simultaneous Quantification of the Enterococcus spp. and the Human Fecal-associated HF183 Marker in Waters

This manuscript describes a duplex digital PCR assay that can be used to simultaneously quantify Enterococcus spp. and the HF183 genetic markers as indicators of general and human-associated fecal contamination in recreational waters.

双工数字PCR方法的同时定量<em&gt;肠球菌。</em&gt;和人的粪便相关HF183标记的水域

This manuscript describes a duplex digital PCR assay (EntHF183 dPCR) for simultaneous quantification of Enterococcus spp. and the human fecal-associated ...

Protocol for Microplastics Sampling on the Sea Surface and Sample Analysis

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of scientific publications address microplastic pollution of the sea surface. The protocol below describes the methodology for sampling, sample preparation, separation and chemical identification of microplastic particles. A manta net fixed on an &#187;A frame&#171; attached to the side of the vessel was used for sampling. Microplastic particles caught in the cod end of the net were separated from samples by visual identification and use of stereomicroscopes. Particles were analyzed for their size using an image analysis program and for their chemical structure using ATR-FTIR and micro FTIR spectroscopy. The described protocol is in line with recommendations for microplastics monitoring published by the Marine Strategy Framework Directive (MSFD) Technical Subgroup on Marine Litter. This written protocol with video guide will support the work of researchers that deal with microplastics monitoring all over the world.

The protocol below describes the methodology for: microplastics sampling on the sea surface, separation of microplastic and chemical identification of particles. This protocol is in line with the recommendations for microplastics monitoring published by the MSFD Technical Subgroup on Marine Litter.

协议塑料微粒采样的海面和样品分析

Microplastic pollution in the marine environment is a scientific topic that has received increasing attention over the last decade. The majority of ...

Ecotoxicological Method with Marine Bacteria Vibrio anguillarum to Evaluate the Acute Toxicity of Environmental Contaminants

Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food webs. Toxicity testing based on microorganisms are widely used because they are relatively quick, reproducible, cheap, and are not associated with ethical issues. Here, we describe an ecotoxicological method to evaluate the biological response of the marine bacterium Vibrio anguillarum. This method assesses the acute toxicity of chemical compounds, including new contaminants such as nanoparticles, as well as environmental samples. The endpoint is the reduction of bacterial culturability (i.e., the capability to replicate and form colonies) due to exposure to a toxicant. This reduction can be generally referred to as mortality. The test allows for the determination of the LC50, the concentration that causes a 50% decrease of bacteria actively replicating and forming colonies, after a 6 h exposure. The culturable bacteria are counted in terms of colony forming units (CFU), and the &#34;mortality&#34; is evaluated and compared to the control. In this work, the toxicity of copper sulphate (CuSO4) was evaluated. A clear dose-response relationship was observed, with a mean LC50 of 1.13 mg/L, after three independent tests. This protocol, compared to existing methods with microorganisms, is applicable in a wider range of salinity and has no limitations for colored/turbid samples. It uses saline solution as the exposure medium, avoiding any possible interferences of growth medium with the investigated contaminants. The LC50 calculation facilitates comparisons with other bioassays commonly applied to ecotoxicological assessments of the marine environment.

Ecotoxicological Method with Marine Bacteria Vibrio anguillarum to Evaluate the Acute Toxicity of Environmental Contaminants

This work describes a new protocol to assess the ecotoxicity of pollutants, including emerging contaminants such as nanomaterials, using the marine bacterium Vibrio anguillarum. This method allows for the determination of the LC50, or mortality, the concentration that causes a 50% decrease in bacteria culturability, after a 6 h exposure.

生物毒理学方法与海洋细菌<em&gt;鳗弧菌</em&gt;评估环境污染物的急性毒性

Bacteria are an important component of the ecosystem, and microbial community alterations can have a significant effect on biogeochemical cycling and food ...

Continuous Instream Monitoring of Nutrients and Sediment in Agricultural Watersheds

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in water resources is a prerequisite for understanding the drivers of the pollutant loads and for making informed water resource management decisions. The commonly used &#34;grab sampling&#34; method provides the concentrations of pollutants at the time of sampling (i.e., a snapshot concentration) and may under- or overpredict the pollutant concentrations and loads. Continuous monitoring of nutrients and sediment has recently received more attention due to advances in computing, sensing technology, and storage devices. This protocol demonstrates the use of sensors, sondes, and instrumentation to continuously monitor in situ nitrate, ammonium, turbidity, pH, conductivity, temperature, and dissolved oxygen (DO) and to calculate the loads from two streams (ditches) in two agricultural watersheds. With the proper calibration, maintenance, and operation of sensors and sondes, good water quality data can be obtained by overcoming challenging conditions such as fouling and debris buildup. The method can also be used in watersheds of various sizes and characterized by agricultural, forested, and/or urban land.

With the advancement of technology and the rise in end-user expectations, the need and use of higher temporal resolution data for pollutant load estimation has increased. This protocol describes a method for continuous in situ water quality monitoring to obtain higher temporal resolution data for informed water resource management decisions.

农业流域营养盐和泥沙的连续河道监测

Pollutant concentrations and loads in watersheds vary considerably with time and space. Accurate and timely information on the magnitude of pollutants in ...

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current measures to combat such pathogens have proved too conservative and, thus, not sufficiently effective, and they can potentially pose environmental risks. Therefore, it is extremely necessary to identify host-resistance factors to assist in controlling plant diseases naturally through the identification of resistant germplasm, the isolation and characterization of resistance genes, and the molecular breeding of resistant cultivars. In this regard, there is need to establish an accurate, rapid, and large-scale inoculation method to breed and develop plant resistance genes. The rice blast fungal pathogen Magnaporthe grisea causes severe disease symptoms and yield losses. Recently, M. grisea has emerged as a model organism for studying the mechanisms of plant-fungal pathogen interactions. Hence, we report the development of a plant virulence test method that is specific for M. grisea. This method provides for both spray inoculation with a conidial suspension and wounding inoculation with mycelium cubes or droplets of conidial suspension. The key step of the wounding inoculation method for detached rice leaves is to make wounds on plant leaves, which avoids any interference caused by host penetration resistance. This spray/wounding protocol contributes to the rapid, accurate, and large-scale screening of the pathotypes of M. grisea isolates. This integrated and systematic plant infection method will serve as an excellent starting point for gaining a broad perspective of issues in plant pathology.

The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea

Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding.

植物感染试验: 以植物病原菌为载体的喷雾和伤口介导接种

Plants possess a powerful system to defend themselves against potential threats by pathogenic fungi. For agriculturally important plants, however, current ...

Construction of a Low-cost Mobile Incubator for Field and Laboratory Use

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water quality. However, commercially available incubators are often costly, difficult to transport, not flexible in terms of volume, and/or poorly adapted to local field conditions where access to electricity is unreliable. The purpose of this study was to develop an adaptable, low-cost and transportable incubator that can be constructed using readily available components. The electronic core of the incubator was first developed. These components were then tested under a range of ambient temperature conditions (3.5 &#176;C - 39 &#176;C) using three types of incubator shells (polystyrene foam box, hard cooler box, and cardboard box covered with a survival blanket). The electronic core showed comparable performance to a standard laboratory incubator in terms of the time required to reach the set temperature, inner temperature stability and spatial dispersion, power consumption, and microbial growth. The incubator set-ups were also effective at moderate and low ambient temperatures (between 3.5 &#176;C and 27 &#176;C), and at high temperatures (39 &#176;C) when the incubator set temperature was higher. This incubator prototype is low-cost (&lt; 300 USD) and adaptable to a variety of materials and volumes. Its demountable structure makes it easy to transport. It can be used in both established laboratories with grid power or in remote settings powered by solar energy or a car battery. It is particularly useful as an equipment option for field laboratories in areas with limited access to resources for water quality monitoring.

This paper describes a method for building an adaptable, low-cost and transportable incubator for microbial testing of drinking water. Our design is based on widely available materials and can operate under a range of field conditions, while still offering the advantages of higher-end laboratory-based models.

建造供野外和实验室使用的低成本移动孵化器

Incubators are essential for a range of culture-based microbial methods, such as membrane filtration followed by cultivation for assessing drinking water ...